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Investigation of Clarithromycin Resistance-Associated Mutations and Virulence Genotypes of Helicobacter pylori Isolated from Iranian Population: A Cross-Sectional Study

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Abstract

Antibiotic resistance has brought into question the efficiency of clarithromycin which is a vital component of eradication therapy for Helicobacter pylori infection. The point mutations within the 23S rRNA sequence of H. pylori isolates which contribute to clarithromycin resistance have yet to be fully characterized. This study was aimed to detect clarithromycin resistance-associated mutations and assess the prevalence of key virulence factors of H. pylori among Iranian patients. Amplification of 16S rRNA and glmM genes were done to identify H. pylori. Minimal inhibitory concentration (MIC) of clarithromycin in 82 H. pylori clinical isolates was determined by agar dilution method. Subsequently, various virulence markers including cagA, vacA, sabA, babA, and dupA of H. pylori were identified by PCR. PCR-sequencing was applied to detect point mutations in the 23S rRNA gene. Based on MIC values, 43.9% of H. pylori isolates showed resistance to clarithromycin. The babA and cagA genes were detected in 92.7% and 82.9% of isolates, assigned to be higher than other virulence factors. No significant relationship was found between the H. pylori virulence genotypes and clarithromycin susceptibility (P > 0.05). Analyzing the 23S rRNA sequences revealed A2143G (4/48, 8.3%) and A2142G (3/48, 6.2%) as the most prevalent mutations in clarithromycin-resistant isolates. Additionally, several novel mutations including G2220T, C2248T, A2624C, G2287A, T2188C, G2710C, C2248T, G2269A, and G2224T were also detected among either resistant or susceptible isolates. Our findings revealed the presence of several point mutations in the 23S rRNA gene of H. pylori isolates which may be associated with resistance to clarithromycin.

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Data Availability

All data generated or analyzed during this study are included in this published article [and its supplementary information files].

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Acknowledgements

The authors would like to thank the sample collection team at Taleghani hospital and the staffs in Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Funding

This study was financially supported by a research grant (no. RIGLD 878) from Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

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Authors and Affiliations

  1. Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Helia Alavifard, Nasrin Mirzaei & Abbas Yadegar

  2. Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Kaveh Baghaei, Amir Sadeghi & Mohammad Reza Zali

  3. School of Medicine & School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin 2, Ireland

    Sinéad Marian Smith

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  1. Helia Alavifard
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Contributions

HA performed the susceptibility testing and PCR assay for H. pylori strains. NM provided the draft of the manuscript. AY worked on concept and design of the study, data analysis and interpretation, and writing of manuscript. AY and KB performed the sequence analysis. AY and SMS critically revised the manuscript. AS and MRZ provided clinical consultation for the study. All authors approved the final version of the manuscript.

Corresponding author

Correspondence to Abbas Yadegar.

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Conflict of interest

The authors have no conflicts of interest to declare.

Ethical Approval

The study protocol was approved by the Institutional Ethical Review Committee of the Research Institute for Gastroenterology and Liver Diseases at Shahid Beheshti University of Medical Sciences (Project No. IR.SBMU.RIGLD.REC.1395.878).

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All participants and/or their legal guardians signed written informed consent before enrollment in the study.

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All authors approved the final version of the manuscript and the authorship list.

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Alavifard, H., Mirzaei, N., Yadegar, A. et al. Investigation of Clarithromycin Resistance-Associated Mutations and Virulence Genotypes of Helicobacter pylori Isolated from Iranian Population: A Cross-Sectional Study. Curr Microbiol 78, 244–254 (2021). https://doi.org/10.1007/s00284-020-02295-7

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